Method for treating hyperuricemia employing an SGLT2 inhibitor and composition containing same

ABSTRACT

Methods are provided for treating hyperuricemia, employing an SGLT2 inhibitor alone, or in combination with a supply of carbohydrate, and/or in combination with an inhibitor of uric acid synthesis. Additionally, pharmaceutical compositions comprising an SGLT2 inhibitor with a supply of carbohydrate or an inhibitor of uric acid synthesis or a supply of carbohydrate and an inhibitor of uric acid synthesis are provided in the instant invention and are provided for use in the inventive methods.

This application is a U.S. National Phase Application under 35 U.S.C.§371 of PCT/US09/44156, filed May 15, 2009, which claims the benefit ofpriority to U.S. provisional application Ser. No. 61/055,352, filed May22, 2008, the disclosure of which is hereby incorporated by reference inits entirety.

FIELD OF THE INVENTION

The invention relates to methods for treating hyperuricemia, comprisingadministering an SGLT2 inhibitor alone or in combination with acarbohydrate and/or an agent that inhibits uric acid synthesis, and tocompositions thereof.

BACKGROUND OF THE INVENTION

Hyperuricemia is a condition of high serum total urate levels. In humansand higher primates, uric acid is the final oxidation product of purinecatabolism. In most other mammals, however, the enzyme uricase furtheroxidizes uric acid to allantoin. In human and higher primates, whichlack the enzyme uricase, purine metabolites such as xanthine andhypoxanthine are oxidized by xanthine oxidase to uric acid. In humanblood, uric acid concentrations between 3.6 mg/dL (˜214 μmol/L) and 8.3mg/dL (˜494 μmol/L) are considered normal by the American MedicalAssociation. The presence of total urates including uric acid in theserum is important and beneficial because these compounds are potentantioxidants. In humans, about half the antioxidant capacity of plasmacomes from total urates including uric acid.

On the other hand, high serum total urate levels, or hyperuricemia, areoften associated with several maladies. For example, high serum totalurate levels can lead to a type of arthritis in the joints known asgout. Gout is a condition created by a build up of monosodium urate oruric acid crystals on the articular cartilage of joints, tendons andsurrounding tissues due to elevated concentrations of total urate levelsin the blood stream. The build up of urate or uric acid on these tissuesprovokes an inflammatory reaction of these tissues. Saturation levels ofuric acid in urine may result in one form of kidney stones when the uricacid or urate crystallizes in the kidney. These uric acid stones areradiolucent and so do not appear on an abdominal x-ray. Therefore, theirpresence must be diagnosed by ultrasound. Some patients with gouteventually develop uric kidney stones.

Additionally, high serum total urate levels are often associated withthe so-called metabolic syndrome, including cardiovascular disease andhypertension. Conventionally, it was believed that high total uratelevels are merely innocuous or could even be beneficial because of theantioxidant activity of uric acid. More recently, however, this view hasbeen challenged. Rather, it has been proposed that total urates are atrue risk factor for cardiovascular disease and hypertension. In a ratanimal model, hyperuricemia resulted in lowering endothelial nitricoxide levels, reducing neuronal nitric oxide synthase in the maculadensa of the kidney, and stimulating the rennin-angiotensin system. Overtime, the rats developed renal microvascular lesions and eventuallyhypertension. Heinig et al. Cleveland Clinic Journal of Medicine, 2006,73:1059-1064. Thus, there is evidence that high serum total urate level,or hyperuricemia, is a risk factor for hypertension.

Hyperuricemia is caused either by accelerated generation of total uratesand uric acid through purine metabolism or by impaired excretion oftotal urates in the urine. Consumption of purine-rich diets is one ofthe causes of hyperuricemia. High levels of fructose in the diet mayalso cause hyperuricemia. Other dietary causes are ingestion of highprotein and fat, and starvation. Starvation results in the bodymetabolizing its own muscle mass for energy, in the process releasingpurines into the bloodstream. Hyperuricemia may lead to renal diseasesand may also exacerbate existing renal conditions.

Conventional chronic, prophylactic treatments of gout or other high uricacid-associated diseases include administering to a patient anuricosuric drug, which augments urinary uric acid excretion, such asprobenecid, sulfinpyrazone, or benzbromarone; and/or an inhibitor ofxanthine oxidase, such as allopurinol, febuxostat, or oxypurinol. Axanthine oxidase inhibitor reduces total urate production in the body.Allopurinol, the most commonly used xanthine oxidase inhibitor, isassociated with side-effects in up to 20% of patients. Therefore, thereremains a need for additional safe and effective treatments forhyperuricemia.

SUMMARY OF THE INVENTION

This invention provides methods and reagents for treating hyperuricemiaby inhibiting a sodium glucose transporter expressed in the kidneycalled SGLT2. SGLT2 is a member of a family of proteins that utilizes anelectrochemical sodium gradient to transport glucose, against the sodiumconcentration gradient inside the cells. Different Na+/Glucosetransporters are found in different tissues: SGLT1 is mainly found inintestinal mucosa in the small intestine and the S3 segment of theproximal tubule of the nephron in the kidney; and SGLT2 is mainly foundin the S1 segment of the proximal tubule of the nephron in the kidney.As set forth herein, SGLT2 inhibitors increase urine excretion ofglucose. This increase in urinary glucose excretion is beneficial as atreatment for hyperuricemia.

In one aspect, the invention provides methods for treating hyperuricemiain a mammal comprising administering to the mammal in need of suchtreatment a therapeutically effective amount of an SGLT2 inhibitor. Inone embodiment, the SGLT2 inhibitor is dapagliflozin (compound I). Inyet another embodiment, the SGLT2 inhibitor compound is dapagliflozinPGS (compound Ia). In certain embodiments of this aspect, the methods ofthe invention for treating hyperuricemia in a mammal compriseadministering to the mammal in need thereof a pharmaceutical compositioncomprising a therapeutically effective amount of SGLT2 inhibitor, andoptionally at least one pharmaceutically-acceptable carrier, excipientor diluent.

In another aspect, the invention provides methods for treatinghyperuricemia in a mammal comprising administering to the mammal in needof such treatment a therapeutically effective amount of an SGLT2inhibitor and a supply of carbohydrate. The carbohydrate can be suppliedprior to, after, or concurrently with the SGLT2 inhibitor and,optionally, another anti-hyperuricemic agent.

In a further aspect, these methods of the invention compriseadministering to the mammal, in addition to a therapeutically effectiveamount of an SGLT2 inhibitor, an inhibitor of uric acid synthesis, suchas a xanthine oxidase inhibitor, where the inhibitor of uric acidsynthesis is administered prior to, after, or concurrently with theSGLT2 inhibitor. In a particular embodiment of the invention, theinhibitor of uric acid synthesis is a xanthine oxidase inhibitor.

In yet another embodiment, the methods of the invention comprisesadministering to the mammal a therapeutically effective amount of anSGLT2 inhibitor, a supply of carbohydrate, and an inhibitor of uric acidsynthesis.

In certain embodiments of this aspect, the methods of the invention fortreating hyperuricemia in a mammal comprise administering to the mammalin need thereof a pharmaceutical composition comprising atherapeutically effective amount of SGLT2 inhibitor, a supply ofcarbohydrate or an inhibitor of uric acid synthesis or both a supply ofcarbohydrate and optionally an inhibitor of uric acid synthesis, and atleast one pharmaceutically-acceptable carrier, excipient or diluent.

In a further aspect, the invention provides pharmaceutical compositionsfor treating hyperuricemia comprising an SGLT2 inhibitor and aninhibitor of uric acid synthesis. In one embodiment, the pharmaceuticalcompositions further comprise at least one pharmaceutically-acceptablecarrier, excipient or diluent.

In another aspect, the invention provides pharmaceutical compositionsfor treating hyperuricemia comprising an SGLT2 inhibitor and a source ofcarbohydrate or an inhibitor of uric acid synthesis or both a source ofcarbohydrate and an inhibitor of uric acid synthesis. In one embodiment,the pharmaceutical compositions further comprise at least onepharmaceutically-acceptable carrier, excipient or diluent.

In yet another aspect, the invention provides methods for treating goutdue to hyperuricemia in a mammal comprising administering to the mammalin need of such treatment a therapeutically effective amount of an SGLT2inhibitor. In one embodiment, the SGLT2 inhibitor is dapagliflozin(compound I). In yet another embodiment, the SGLT2 inhibitor compound isdapagliflozin PGS (compound Ia). In certain embodiments of this aspect,the methods of the invention for treating hyperuricemia in a mammalcomprise administering to the mammal in need thereof a pharmaceuticalcomposition comprising a therapeutically effective amount of SGLT2inhibitor, and optionally at least one pharmaceutically-acceptablecarrier, excipient or diluent.

In another aspect, the invention provides methods for treating gout in amammal comprising administering to the mammal in need of such treatmenta therapeutically effective amount of an SGLT2 inhibitor and a supply ofcarbohydrate. The carbohydrate can be supplied prior to, after, orconcurrently with the SGLT2 inhibitor. In a further aspect, thesemethods of the invention comprise administering to the mammal, inaddition to a therapeutically effective amount of an SGLT2 inhibitor, aninhibitor of uric acid synthesis where the inhibitor of uric acidsynthesis is administered prior to, after, or concurrently with theSGLT2 inhibitor. In a particular embodiment of the invention, theinhibitor of uric acid synthesis is a xanthine oxidase inhibitor. In yetanother embodiment, the methods of the invention comprises administeringto the mammal a therapeutically effective amount of an SGLT2 inhibitor,a supply of carbohydrate, and an inhibitor of uric acid synthesis. Incertain embodiments of this aspect, the methods of the invention fortreating gout in a mammal comprise administering to the mammal in needthereof a pharmaceutical composition comprising a therapeuticallyeffective amount of SGLT2 inhibitor, a supply of carbohydrate or aninhibitor of uric acid synthesis or both a supply of carbohydrate and aninhibitor of uric acid synthesis, and at least onepharmaceutically-acceptable carrier, excipient or diluent.

In another aspect, the invention provides pharmaceutical compositionsfor treating hyperuricemia comprising an SGLT2 inhibitor and a source ofcarbohydrate or an inhibitor of uric acid synthesis or both a source ofcarbohydrate and an inhibitor of uric acid synthesis. In one embodiment,the pharmaceutical compositions further comprise at least onepharmaceutically-acceptable carrier, excipient or diluent.

Specific preferred embodiments of the present invention will becomeevident from the following more detailed description of certainpreferred embodiments and the claims.

BRIEF DESCRIPTION OF THE FIGURES

The invention is illustrated by reference to the accompanying drawingsdescribed below.

FIG. 1 shows two bar graphs comparing urine glucose excretion in healthyindividuals that were treated with GSK 869,682 (top panel) anddapagliflozin PGS (bottom panel).

FIG. 2 shows a graph depicting urine glucose excretion in diabeticsubjects treated with different doses of dapagliflozin PGS.

FIG. 3 shows a bar graph depicting urine glucose excretion in diabeticsubjects treated with different doses of dapagliflozin PGS.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and reagents directed towards treatinghyperuricemia in a mammal in need thereof. In particular, these methodsand reagents are capable of facilitating the lowering of serum totalurate levels in the animal, and alleviation of the symptoms of gout,hypertension, renal failure, and other hyperuricemia-associatedconditions in the mammal.

In one aspect, the invention provides methods for treating hyperuricemiain a mammal comprising administering to the mammal in need of suchtreatment a therapeutically effective amount of a sodium glucosetransporter 2 (SGLT2) inhibitor. In another aspect, the inventionprovides methods for treating hyperuricemia in a mammal furthercomprising administering to the mammal at least one additional reagentthat facilitates and enhances the effects of the SGLT2 inhibitor on thetreatment of hyperuricemia. In certain embodiments, the methods of theinvention for treating hyperuricemia further comprise administering tothe mammal in need thereof a supply of carbohydrate. In certainembodiments of this aspect, the methods of the invention for treatinghyperuricemia in a mammal comprise administering to the mammal in needthereof a pharmaceutical composition comprising a therapeuticallyeffective amount of SGLT2 inhibitor, optionally a supply ofcarbohydrate, and further optionally at least onepharmaceutically-acceptable carrier, excipient or diluent.

In certain embodiments, the mammal is a human. In yet other embodiments,the mammal suffers from gout and/or renal failure as a result of highserum uric acid levels. The present invention provides methods forlowering serum total urate and treating gout, renal failure and otherhyperuricemia-associated diseases.

In another aspect, the invention provides methods for treatinghyperuricemia in a mammal comprising administering to the mammal in needof such treatment a therapeutically effective amount of a sodium glucosetransporter 2 (SGLT2) inhibitor and an inhibitor of uric acid synthesiswherein the inhibitor of uric acid synthesis is administered before,after, or concurrently with the SGLT2 inhibitor. In certain embodiments,the invention provides methods for treating hyperuricemia in a mammalcomprising administering to the mammal in need of such treatment atherapeutically effective amount of a SGLT2 inhibitor, a supply ofcarbohydrate, and an inhibitor of uric acid synthesis.

In certain embodiments of this aspect, the methods of the invention fortreating hyperuricemia in a mammal comprise administering to the mammalin need thereof a pharmaceutical composition comprising atherapeutically effective amount of SGLT2 inhibitor and a supply ofcarbohydrate or an inhibitor of uric acid synthesis or both acarbohydrate and an inhibitor of uric acid synthesis, and optionally atleast one pharmaceutically-acceptable carrier, excipient or diluent. Inone embodiment, the inhibitor of uric acid synthesis is a xanthineoxidase inhibitor.

In yet another aspect, the invention provides a SGLT2 inhibitor for usein therapy in treating hyperuricemia. In a further aspect, the inventionprovides the use of a SGLT2 inhibitor in the manufacture of a medicamentfor treating hyperuricemia. In a further aspect, the invention providesa combination of a SGLT2 inhibitor and a supply of carbohydrate or aninhibitor of uric acid synthesis or a supply of carbohydrate and aninhibitor of uric acid synthesis as a medicament for the treatment ofhyperuricemia. In another aspect, the invention provides use of a SGLT2inhibitor in the manufacture of a medicament for treating hyperuricemia,in which such treatment or prevention comprises a combination with asupply of carbohydrate or an inhibitor of uric acid synthesis or asupply of carbohydrate and an inhibitor of uric acid synthesis, forconcurrent or sequential use. In certain embodiments, the supply ofcarbohydrate and/or the inhibitor of uric acid synthesis is usedsequentially, either before or after the treatment with the SGLT2inhibitor. In certain embodiments, the inhibitor of uric acid synthesisis a xanthine oxidase inhibitor.

Inhibitors of uric acid synthesis suitable for use in the currentinvention include, but are not limited to, xanthine oxidase inhibitors.Examples of xanthine oxidase inhibitors include, without limitation,allopurinol, febuxostat and oxypurinol.

The term “a supply of carbohydrate,” or “a source of carbohydrate” asused herein refers to simple or complex carbohydrate either from thedaily intake of carbohydrate in the diet or from a supplement formulatedwith the SGLT2 inhibitor of the present invention or provided inconjunction with the SGLT2 inhibitor. A source of carbohydrate can beused advantageously with the SGLT2 inhibitor of the present invention toincrease urine volume. Non-limiting examples of carbohydrate includemonosaccharide such as glucose, disaccharides such as sucrose, andcomplex oligosaccharides or polysaccharides such as starch. The supplyof carbohydrate can be provided to the mammal in need of thehyperuricemia treatment before, after, or concurrent with the SGLT2inhibitor. The carbohydrate of the current invention can be supplied inthe range of about 30 to about 270 g per day, preferably in the range ofabout 60 to about 180 g per day, and most preferably about 90 g, orclose to about one third of the mammal's daily carbohydrate intake, perday. It is within the knowledge of one skilled artisan or physician todetermine the suitable amount of carbohydrate to be used in the presentinvention.

The methods of this invention can be used to treat hyperuricemia innon-diabetic as well as diabetic patients. In one embodiment, the SGLT2inhibitors employed in the above-defined methods of the invention willnot cause hypoglycemia in a non-diabetic mammal, i.e. a mammal that doesnot suffer from hyperglycemia. In another embodiment, the SGLT2inhibitors employed in the above-defined methods of the invention willnot cause hypoglycemia in a diabetic mammal. It will be understood bythe skilled worker that administered amounts and in vivo concentrationsof the SGLT2 inhibitors used according to the methods of the inventioncan be chosen to have anti-hyperuricemia effects without disturbing therecipient's plasma glucose homeostasis.

Thus, in carrying out the methods of the invention for treatinghyperuricemia, the SGLT2 inhibitor can be administered to a patient inneed of such treatment in a dose which can be as high as that used totreat hyperuricemia but less than an amount which could causehypoglycemia. The daily dose can be lowered as successful treatment ofhyperuricemia is achieved. For example, depending upon the patient, andthe specific SGLT2 inhibitor employed, the SGLT2 inhibitor can be orallyadministered in a hyperuricemia treating amount from about 1 to about1000 mg per day, preferably from about 2 to about 400 mg/day, preferably2.5 to about 75 mg/day, and more preferably 20 to about 50 mg/day, whichcan be administered in a single dose or in the form of individual dosesfrom 1 to 4 times per day.

In certain embodiments of the methods of the invention, the SGLT2inhibitor is advantageously administered in conjunction with a supply ofcarbohydrate to a mammal in need of a hyperuricemia treatment. Withoutbeing limited to particular mechanisms, the combination of an SGLT2inhibitor and a supply of carbohydrate can further promote osmoticexcretion of glucose. Without being limited to a particular mechanism,the beneficial effects of higher local concentration of glucose in thekidney due to the effects of the SGLT2 inhibitor and further enhanced bythe supply of carbohydrate may be two fold: the elevated localconcentration of glucose in the kidney may prevent the re-adsorption ofuric acid back into the blood stream, and the increased osmoticexcretion of glucose increases water release in the urine, whichfacilitates the excretion of excess uric acid into the urine. Theinvention also provides pharmaceutical compositions or pharmaceuticalcombinations for treating hyperuricemia comprising a therapeuticallyeffective amount of an SGLT2 inhibitor. In yet another aspect, theinvention provides pharmaceutical compositions for treatinghyperuricemia comprising a therapeutically effective amount of an SGLT2inhibitor and an inhibitor of uric acid synthesis. Advantageously, thepharmaceutical composition may further comprise a supply ofcarbohydrate. In certain embodiments of this aspect, the SGLT2 inhibitoris dapagliflozin. In further embodiments, the SGLT2 inhibitor isdapagliflozin PGS. In certain other embodiments, the SGLT2 inhibitor isremogliflozin, remogliflozin etabonate, sergliflozin or sergliflozinetabonate.

Other SGLT2 inhibitors suitable for use in this aspect of the inventionare described throughout the specification, including without limitationAVE2268, TS-033, YM-543, BI 10773, BI 44847, and TA-7284. In certainembodiments, the pharmaceutical compositions further comprise at leastone pharmaceutically-acceptable carrier, excipient or diluent. Theinvention provides methods for treating hyperuricemia comprisingadministering to a mammal in need thereof a pharmaceutical compositionof the invention.

The SGLT2 inhibitors suitable for use in accordance with the inventionalso can be administered by injection to a patient in a hyperuricemiatreating amount from about 1 to about 100 mg/day, preferably from about1 to about 30 mg/day.

The SGLT2 inhibitors employed in the invention are most preferablyselective for SGLT2 relative to SGLT-1. High selectivity for SGLT2, asis the case with dapagliflozin, is advantageous for use in the currentinvention because it avoids the unpredictable effects of intestinalSGLT1 inhibition.

Selectivity for SGLT2 of a given inhibitor can be determined bycomparing the EC₅₀ values measured in the SGLT1 and SGLT2 assay.Briefly, Human SGLT1 (hSGLT1) and human SGLT2 (hSGLT2) full-length cDNAsequences were cloned by PCR using MARATHON READY™ human kidney cDNA(Clontech, Mountain View, Calif.), with primers designed from publishedsequences (Genbank accession numbers NM_(—)003041 and NM_(—)000343). ThehSGLT1 and hSGLT2 sequences were cloned into pIRESneo vector (Clontech,Mountain View, Calif.) for mammalian expression and were stablytransfected into Chinese hamster ovary (CHO) cells. SGLT-expressingclones were selected based on resistance to G418 antibiotic (GENETICIN®,Invitrogen, Carlsbad, Calif.) and activity in the¹⁴C-α-methyl-D-glucopyranoside (¹⁴C-AMG) uptake assay.

Cells expressing hSGLT1 or hSGLT2 were maintained using standard cellculture techniques. Assays for sodium-dependent glucose transport in96-well plates were initiated by adding 100 μl/well of protein-freeassay buffer containing sodium (Hepes/Tris pH 7.4, 137 mM NaCl, 5.4 mMKCl, 2.8 mM CaCl₂, 1.2 mM MgSO₄), 10 μM ¹⁴C-AMG and inhibitor ordimethyl sulfoxide (DMSO) vehicle, and plates were incubated for 2 h at37° C. Sodium-dependent ¹⁴C-AMG uptake was calculated by subtracting thecounts per minute (CPM) observed under sodium-free uptake conditionsfrom the counts observed under sodium-containing conditions. Inhibitorswere assayed at various concentrations in triplicate in the presence ofsodium, and the percent inhibition was calculated by comparing CPM ininhibitor-containing wells with CPM in wells containing only DMSOvehicle. Phlorizin, a known SGLT inhibitor, was evaluated in parallel inevery assay. A dose-response curve was fitted to an empiricalfour-parameter model using XL Fit (IDBS, Guilford, UK) to determine theinhibitor concentration at half-maximal response (EC₅₀). SGLT2selectivity is represented as a ratio of EC₅₀ in favor of SGLT2. AnSGLT2 inhibitor with an EC₅₀ selective ratio of at least 10, and morepreferably at least 100, in favor of SGLT2 is suitable for use in theinstant invention.

The SGLT2 inhibitors suitable for use in accordance with the inventioncomprise C-arylglucosides or O-arylglucosides. SGLT2 inhibitorsC-arylglucosides and O-arylglucosides are effective in treatingdiabetes. See U.S. Pat. No. 6,774,112, which is incorporated herein byreference in its entirety. As disclosed herein, it was unexpectedlydiscovered that this class of SGLT2 inhibitors not only can increaseglucose excretion in urine, but can also increase urine volume. Urinaryexcretion of glucose requires concomitant excretion of electrolyte-freewater so that the final outcome produces a diuresis without releasingsodium to the urine. Thus, the methods of the invention using an SGLT2inhibitor provide an effective treatment for hyperuricemia partlybecause it promotes a gradual release of electrolyte-free water in theprocess of glucose excretion in the urine. To the Applicant's knowledge,no other known anti-diabetic drugs in humans can cause glucosuria.Advantageously and unexpectedly, this class of SGLT2 inhibitors of thecurrent invention increases urine volume over time. Without beinglimited to particular mechanisms, the increased urine volume mayfacilitate release of excess uric acid in the blood stream into urine.In certain embodiment of the invention, the SGLT2 inhibitor is a C-arylglucoside.

Examples of C-arylglucoside (also referred to as C-glucosides) SGLT2inhibitors which can be employed in the method of the invention,include, but are not limited to the following:

1) C-aryl glucosides as disclosed in U.S. Pat. Nos. 6,515,117 and6,414,126, the disclosures of which are incorporated herein by referencein their entirety for any purpose

2) C-aryl glucosides as described in U.S. patent application Ser. No.11/233,617 (U.S. Patent Application Publication No. 2006/0063722 A1),the disclosure of which is incorporated herein by reference in itsentirety;

3) C-aryl glucosides described in U.S. Pat. No. 6,774,112, thedisclosure of which is incorporated herein by reference in its entirety;

4) Glucopyranosyl-substituted benzene derivatives as disclosed in U.S.Patent Application Publication No. 2005/0209166, the disclosure of whichis incorporated herein by reference in its entirety;

5) D-pyranosyl-substituted phenyl compounds as disclosed in U.S. PatentApplication Publication No. S 2006/0074031, the disclosure of which isincorporated herein by reference in its entirety;

6) D-xylopyranosyl-substituted compounds as disclosed in U.S. PatentApplication Publication No. 2006/0035841, the disclosure of which isincorporated herein by reference in its entirety;

7) D-xylopyranosyl-substituted phenyl compounds as disclosed in U.S.Patent Application Publication No. 2006/0009400, the disclosure of whichis incorporated herein by reference in its entirety;

8) D-glucopyranosyl-phenyl-substituted compounds as disclosed in U.S.Patent Application Publication No. 2006/0025349, the disclosure of whichis incorporated herein by reference in its entirety;

9) C-glycoside derivatives as disclosed in U.S. Patent ApplicationPublication No. 2006/0122126, the disclosure of which is incorporatedherein by reference in its entirety;

10) D-xylopyranosyl-substituted phenyl compounds as disclosed in U.S.Patent Application Publication No. 2006/0019948, the disclosure of whichis incorporated herein by reference in its entirety;

Examples of O-glucoside SGLT2 inhibitors which can be employed in themethods and pharmaceutical compositions of the invention include, butare not limited to those described in the the following:

1) 5-Thio-β-D-glucopyranoside as disclosed in U.S. Patent ApplicationPublication No. 2006/0194809, the disclosure of which is incorporated byreference in its entirety for any purpose:

2) Glucopyranyloxybenzene derivatives of as disclosed in WO 03/01180,the disclosure of which is incorporated by reference in its entirety forany purpose:

3) Pyrazole derivatives as disclosed in U.S. Pat. No. 6,908,905, thedisclosure of which is incorporated herein by reference for any purpose:

4) Pyrazole compounds as disclosed in U.S. Pat. No. 6,815,428, thedisclosure of which is incorporated herein by reference for any purpose:

5) O-glucosylated benzamide compounds as disclosed in U.S. Pat. No.6,555,519, the disclosure of which is incorporated herein by referencein its entirety for any purpose:

6) O-arylglucoside (or O-glucoside) compounds as disclosed in U.S. Pat.No. 6,683,056, the disclosure of which is incorporated herein byreference in its entirety for any purpose:

Other O-aryl glucosides SGLT2 inhibitors which can be used in theinvention are disclosed in the following references, each of which isincorporated herein by reference in its entirety for any purpose. EP598359A1, EP 0850948A1, JP 09188625A, JP 09124685A, JP 09124684, EP773226-A1, and JP 08027006-A, EP 684254-A1.

Other disclosures and publications disclosing SGLT2 inhibitors that canbe employed in the methods and pharmaceutical compositions of theinvention are as follows: K. Tsujihara et al., Chem. Pharm. Bull.,44:1174-1180 (1996); M. Hongu et al., Chem. Pharm. Bull., 46:22-33(1998); M. Hongu et al., Chem. Pharm. Bull., 46:1545-1555 (1998); and A.Oku et al., Diabetes, 48:1794-1800 (1999) and. JP 10245391 (Dainippon).

Preferred SGLT2 inhibitors that can be employed in the present inventioninclude dapagliflozin, remigliflozin, antisense oligonucleotide ISIS388626, sergliflozin and those disclosed in U.S. Patent ApplicationPublication No. 2005/0233982 (Boehringer Ingelheim Corp.), U.S. PatentApplication Publication No. 2005/0119192 (Kissei Pharmaceutical Co.), WO2006/035796 (Kissei Pharmaceutical Co.), JP 2006/117651 (TaishoPharmaceutical Co.), JP 2004/4359630 (Yamanouchi Pharmaceutical Co.), WO2006/080421 (Chugai Seiyaku Kabushiki Kaishi), U.S. Patent ApplicationPublication No. 2005/0233988 (Tanabe Seiyaku Co.), WO 2005/012321(Tanabe Seiyalcu Co.), U.S. Pat. No. 7,015,201 (Ajinomoto Co.), WO2006/058597 (Merck Patent GmbH), WO 2006/011469 (Chugai SeiyakuKabushiki Kaisha), US 2003/0195235 (Johnson & Johnson), and WO2006/037537 (Boehringer Ingelheim), the disclosure of each of which isherein incorporated by reference in its entirety for any purpose.

In a preferred aspect, the invention provides SGLT2 inhibitors for usein the methods and pharmaceutical compositions of the invention that aredisclosed in U.S. Pat. Nos. 6,414,126 and 6,515,117, more preferably theSGLT2 inhibitor is compound I or dapagliflozin

or a pharmaceutically acceptable salt thereof, all stereoisomersthereof, or a prodrug ester thereof.

In another preferred aspect, the invention provides an SGLT2 inhibitorfor use in the methods and pharmaceutical compositions of the inventionthat is compound IIIA

or a pharmaceutically acceptable salt thereof, all stereoisomersthereof, or a prodrug ester thereof.

In another preferred aspect, the invention provides an SGLT2 inhibitorfor use in the methods and pharmaceutical compositions of the inventionthat is compound II

or a pharmaceutically acceptable salt thereof, all stereoisomersthereof, or a prodrug ester thereof.

In another preferred aspect, the invention provides crystalline forms ofcompound I including the crystalline forms disclosed in U.S. applicationSer. No. 11/765,481, the disclosure of which is incorporated herein byreference in its entirety for any purpose. A most preferred crystallineform for use in the methods and pharmaceutical compositions of theinvention is the (S)-propylene glycol solvate of the compound of formulaI, namely Compound Ia or dapagliflozin PGS

Compound Ia or dapagliflozin PGS is prepared as described in U.S.application Ser. No. 11/765,481, published as U.S. Patent ApplicationPublication No. 2008/0004336, which is incorporated by reference herein.

The SGLT2 inhibitor employed in accordance with the invention can beadministered to various mammalian species, such as dogs, cats, cattle,humans, etc., in need of treatment. These agents can be administeredsystemically, such as orally or parenterally.

The invention also provides pharmaceutical compositions that comprisecompounds of the invention formulated together with one or morepharmaceutically acceptable carriers. The pharmaceutical compositionscan be specially formulated for oral administration in solid or liquidform, for parenteral injection or for rectal administration.

The pharmaceutical compositions of this invention can be administered tohumans and other mammals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, bucally or as anoral or nasal spray. The term “parenterally,” as used herein, refers tomodes of administration which include intravenous, intramuscular,intraperitoneal, intrasternal, subcutaneous and intraarticular injectionand infusion.

The SGLT-2 inhibitor can be incorporated in a conventional systemicdosage form, such as a tablet, capsule, elixir or injectableformulation. The above dosage forms will also include the necessaryphysiologically acceptable carrier material, excipient, lubricant,buffer, antibacterial, bulking agent (such as mannitol), antioxidants(ascorbic acid or sodium bisulfite) or the like. Oral dosage forms arepreferred, although parenteral forms are quite satisfactory as well.

Pharmaceutical compositions of this invention for parenteral injectioncomprise pharmaceutically acceptable sterile aqueous or nonaqueoussolutions, dispersions, suspensions or emulsions as well as sterilepowders for reconstitution into sterile injectable solutions ordispersions just prior to use. Examples of suitable aqueous andnonaqueous carriers, diluents, solvents or vehicles include water,ethanol, polyols (such as glycerol, propylene glycol, polyethyleneglycol and the like), vegetable oils (such as olive oil), injectableorganic esters (such as ethyl oleate) and suitable mixtures thereof.Proper fluidity can be maintained, for example, by the use of coatingmaterials such as lecithin, by the maintenance of the required particlesize in the case of dispersions and by the use of surfactants.

These compositions may also contain adjuvants such as preservative,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms can be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid and the like. It may also be desirableto include isotonic agents such as sugars, sodium chloride and the like.Prolonged absorption of the injectable pharmaceutical form can bebrought about by the inclusion of agents which delay absorption such asaluminum monostearate and gelatin.

In some cases, in order to prolong the effect of the drug, it isdesirable to slow the absorption of the drug from subcutaneous orintramuscular injection. This can be accomplished by the use of a liquidsuspension of crystalline or amorphous material with poor watersolubility. The rate of absorption of the drug then depends upon itsrate of dissolution which, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered drug form is accomplished by dissolving or suspending thedrug in an oil vehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe drug in biodegradable polymers such as polylactide-polyglycolide.Depending upon the ratio of drug to polymer and the nature of theparticular polymer employed, the rate of drug release can be controlled.Examples of other biodegradable polymers include poly(orthoesters) andpoly(anhydrides). Depot injectable formulations are also prepared byentrapping the drug in liposomes or microemulsions which are compatiblewith body tissues.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium just prior to use.

Solid dosage forms for oral administration include capsules, tablets,pills, powders and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier, such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol and silicic acid; b) binders such ascarboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone,sucrose and acacia; c) humectants such as glycerol; d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates and sodium carbonate; e) solutionretarding agents such as paraffin; f) absorption accelerators such asquaternary ammonium compounds; g) wetting agents such as cetyl alcoholand glycerol monostearate; h) absorbents such as kaolin and bentoniteclay and i) lubricants such as talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate and mixturesthereof. In the case of capsules, tablets and pills, the dosage form mayalso comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills and granulescan be prepared with coatings and shells such as enteric coatings andother coatings well-known in the pharmaceutical formulating art. Theymay optionally contain opacifying agents and may also be of acomposition such that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

The active compounds can also be in micro-encapsulated form, ifappropriate, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups and elixirs. Inaddition to the active compounds, the liquid dosage forms may containinert diluents commonly used in the art such as, for example, water orother solvents, solubilizing agents and emulsifiers such as ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,dimethyl formamide, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor and sesame oils), glycerol, tetrahydrofurfurylalcohol, polyethylene glycols and fatty acid esters of sorbitan andmixtures thereof.

Besides inert diluents, the oral compositions may also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar, tragacanth and mixtures thereof.

The dose administered is adjusted according to the age, weight, andcondition of the patient, as well as the route of administration, dosageform and regimen, and the desired result. In general, the dosage formsdescribed above can be administered containing amounts of SGLT-2inhibitor of from about 1 to about 1000 mg per day preferably from about2 to about 400 mg per day, in single or divided doses of one to fourtimes daily.

Unless otherwise indicated, dosages and formulations for SGLT2inhibitors used in the methods set forth herein are disclosed in thevarious patents and applications discussed throughout the application,which are incorporated herein in their entireties.

The various formulations of the invention can optionally include one ormore fillers or excipients in an amount within the range of from about 0to about 90% by weight and preferably from about 1 to about 80% byweight such as lactose, sugar, corn starch, modified corn starch,mannitol, sorbitol, inorganic salts such as calcium carbonate and/orcellulose derivatives such as wood cellulose and microcrystallinecellulose.

One or more binders can be present in addition to or in lieu of thefillers in an amount within the range of from about 0 to about 35% andpreferably from about 0.5 to about 30% by weight of the composition.Examples of such binders suitable for use herein includepolyvinylpyrrolidone (molecular weight ranging from about 5000 to about80,000 and preferably about 40,000), lactose, starches such as cornstarch, modified corn starch, sugars, gum acacia and the like as well asa wax binder in finely powdered form (less than 500 microns) such ascarnauba wax, paraffin, spermaceti, polyethylenes or microcrystallinewax.

Where the composition is in the form of a tablet, it may include one ormore tabletting lubricants in an amount within the range of from about0.2 to about 8% and preferably from about 0.5 to about 2% by weight ofthe composition, such as magnesium stearate, stearic acid, palmiticacid, calcium stearate, talc, carnauba wax and the like. Otherconventional ingredients that can optionally be present includepreservatives, stabilizers, anti-adherents or silica flow conditionersor glidants, such as Syloid brand silicon dioxide as well as FD&Ccolors.

Tablets of the invention can also include a coating layer that cancomprise from 0 to about 15% by weight of the tablet composition. Thecoating layer can comprise any conventional coating formulations andinclude one or more film-formers or binders, such as a hydrophilicpolymer like hydroxypropylmethylcellulose, and/or a hydrophobic polymerlike methacrylic acid esters neutral polymer, ethyl cellulose, celluloseacetate, polyvinyl alcohol-maleic anhydride copolymers, β-pinenepolymers, glyceryl esters of wood resins and the like and one or moreplasticizers, such as triethyl citrate, diethyl phthalate, propyleneglycol, glycerin, butyl phthalate, castor oil and the like. Both coretablets as well as coating formulations can contain aluminum lakes toprovide color.

The film formers are applied from a solvent system containing one ormore solvents including water, alcohols like methyl alcohol, ethylalcohol or isopropyl alcohol, ketones like acetone, or ethylmethylketone, chlorinated hydrocarbons like methylene chloride,dichloroethane, and 1,1,1-trichloroethane.

Where a color is employed, the color is applied together with the filmformer, plasticizer and solvent compositions.

It will be recognized by one of skill in the art that the amount of drugrequired for therapeutic effect on administration will, of course, varywith the agent chosen, the nature and severity of the condition and themammal undergoing treatment, and is ultimately at the discretion of thephysician. Furthermore, the optimal quantity and spacing of individualdosages of a drug will be determined by the nature and extent of thetherapeutic effects desired, the form, route and site of administration,the particular patient being treated and that such optima can bedetermined by conventional techniques. It will also be appreciated thatthe optimal course of treatment, for example, the number of doses given,can be ascertained by those skilled in the art using conventional courseof treatment determination tests.

Preferred tablet and capsule formulations in accordance with theinvention are set out below in Table 2.

TABLE 2 Tablet and Capsule Formulations Range Preferred Range Material%/mg by weight of 200 mg %/mg by weight of 200 mg Tablet tablet tabletDapagliflozin 0.1 to 70%/0.2 to 140 mg 1 to 50%/2 to 100 mg BulkingAgent 2 to 95%/4 to 190 mg 10 to 85%/20 to 170 mg Lactose 0 to 95%/0 to190 mg 20 to 75%/10 to 100 mg Microcrystalline cellulose 0 to 95%/0 to190 mg 20 to 75%/40 to 150 mg Disintegrant 0 to 20%/0 to 40 mg 0.25 to10%/0.5 to 20 mg Croscarmellose sodium 0 to 20%/0 to 40 mg 2 to 10%/4 to20 mg Crospovidone 4 to 12%/4 to 20 mg 6 to 10%/12 to 20 mg Lubricant0.1 to 5%/0.2 to 10 mg 0.2 to 2%/0.4 to 4 mg Magnesium Stearate 0.1 to5%/0.2 to 10 mg 0.2 to 2%/0.4 to 4 mg Anti adherent/glidant 0 to 10%/0to 20 mg 1 to 4%/2 to 8 mg Talc, silicon dioxide %/mg by weight of 200mg %/mg by weight of 200 mg Outer Protective Coating Layer tablet tabletCoating polymer, and optional 0.5 to 50%/1 to 100 mg 1 to 5%/2 to 10 mgplasticizer, glidants and color

Preferred stock granulation formulations (for use in capsules) inaccordance with the invention are set out below in Table 3.

TABLE 3 Range Preferred Range Material %/mg by weight of 200 mg %/mg byweight of 200 mg Tablet tablet tablet Dapagliflozin 0.1 to 70%/0.2 to140 mg 1 to 50%/2 to 100 mg Bulking Agent/Binder 2 to 95%/4 to 190 mg 10to 85%/20 to 170 mg Microcrystalline cellulose 1 to 95%/1 to 190 mg 20to 75%/10 to 100 mg Pregelatinized starch 0 to 95%/0 to 190 mg 20 to75%/40 to 150 mg Disintegrant 0 to 20%/0 to 40 mg 0.25 to 10%/0.5 to 20mg Sodium Starch glycolate 0 to 20%/0 to 40 mg 2 to 10%/4 to 20 mgLubricant 0.1 to 5%/0.2 to 10 mg 0.2 to 2%/0.4 to 4 mg MagnesiumStearate 0.1 to 5%/0.2 to 10 mg 0.2 to 2%/0.4 to 4 mg Antiadherent/glidant 0 to 10%/0 to 20 mg 1 to 4%/2 to 8 mg Talc silicondioxide

A typical injectable preparation is produced by aseptically placing 50mg of compounds of the present invention into a vial, asepticallyfreeze-drying and sealing. For use, the contents of the vial are mixedwith 2 mL of physiological saline, to produce an injectable preparation.

SGLT2 inhibitor activity of the compounds of the invention can bedetermined by use of an assay system as set out below.

Assay for SGLT2 Activity

The mRNA sequence for human SGLT2 was cloned by reverse-transcriptionand amplification from human kidney mRNA, using standard molecularbiology techniques. The cDNA sequence was stably transfected into CHOcells, and clones were assayed for SGLT2 activity essentially asdescribed in Ryan et al., 1994, “HK-2: an immortalized proximal tubuleepithelial cell line from normal adult human kidney,” KidneyInternational 45:48-57. Evaluation of inhibition of SGLT2 activity in aclonally selected cell line was performed essentially as described inRyan et al., with the following modifications. Cells were grown in96-well plates for 2-4 days to 75,000 or 30,000 cells per well in F-12nutrient mixture (Ham's F-12; GIBCO, Long Island, N.Y.), 10% fetalbovine serum, 300 μg/ml Geneticin and penicillin-streptomycin. Atconfluence, cells were washed twice with 10 mM Hepes/Tris, pH 7.4, 137mM N-methyl-D-glucamine, 5.4 mM KCl, 2.8 mM CaCl₂, 1.2 mM MgSO₄. Cellsthen were incubated with 10 μM [¹⁴C]AMG, and 10 μM inhibitor (finalDMSO=0.5%) in 10 mM Hepes/Tris, pH 7.4, 137 mM NaCl, 5.4 mM KCl, 2.8 mMCaCl₂, 1.2 mM MgSO₄ at 37° C. for 1.5 hr. Uptake assays were quenchedwith ice cold 1× PBS containing 0.5 mM phlorizin, and cells were thenlysed with 0.1% NaOH. After addition of MicroScint scintillation fluid,the cells were allowed to shake for 1 hour, and then [¹⁴C]AMG wasquantified on a TopCount scintillation counter. Controls were performedwith and without NaCl. For determination of EC₅₀ values, 10 inhibitorconcentrations were used over 2 log intervals in the appropriateresponse range, and triplicate plates were averaged across plates. Ryanet al., Id.

EXAMPLES

The following working Examples are illustrative of the invention. Alltemperatures are expressed in degrees Centigrade unless otherwiseindicated.

Examples 1 to 3

Capsules containing the SGLT2 inhibitor of Formula I (dapagliflozin)were prepared in strengths of 2.5 mg (Example 1), 10 mg (Example 2) and100 mg (Example 3) (as the non-solvated form) as two-piece, gray opaquesize #0 (2.5 mg and 10 mg) and size #00 (for 0 mg) hard gelatincapsules.

Examples 1 and 2

Composition: 25.0 mg of Granulation containing dapagliflozin forCapsules (10.0% w/w as the non-solvated form), filled in Gray, Opaque,Size #0 Capsule Shell.

A. Stock Granulation Composition

Ingredient Amount (% w/w) Dapagliflozin PGS¹ 10.0 Pregelatinized Starch,NF 15.0 Microcrystalline Cellulose, NF² 68.75 Sodium Starch Glycolate,NF 3.0 Silicon Dioxide, NF 2.0 Magnesium Stearate, NF³ 1.25 ¹This amountis expressed in terms of the amount of the dapagliflozin at 100% purity.The exact amount will vary depending on the purity of the dapagliflozin.²The amount of microcrystalline cellulose used will vary depending onthe purity of the dapagliflozin. ³The preferred amount is 1.25% (w/w).The range is 1.25-1.50% (w/w).

The stock granulation of Part A and the Example 1 and Example 2 capsuleswere prepared according to the following procedures.

Example 1 B. Example 1 Stock Granulation Procedure

1. Screen dapagliflozin.

2. Screen silicon dioxide.

3. Mix silicon dioxide with dapagliflozin in a suitable blender.

4. Screen pregelatinized starch and microcrystalline cellulose, ifnecessary.

5. Add ingredients from Step 4 to a suitable blender.

6. Add mixture from Step 3 to the blend from Step 5, and mix.

7. Screen sodium starch glycolate.

8. Add ingredient from Step 7 to the blend from Step 6, and mix.

9. Screen the blend from Step 8, and mix.

10. Screen portion of magnesium stearate.

11. Add ingredient from Step 10 to the blend from Step 9, and mix.

12. Densify the blend from Step 11.

13. Reduce the densified blend Step 12.

14. Screen the remaining portion of magnesium stearate.

15. Add ingredient from Step 14 to the granulation from Step 13, andmix.

C. Example 1 Product: Dapagliflozin Capsule, 2.5 mg (as the Non-SolvatedForm)

1. Fill empty capsule shells with sufficient Example 1 Part A stockgranulation for capsules (10.0%) w/w (as the non-solvated form), toprovide 2.5 mg capsules.

2. De-dust the capsules.

Example 2 Product: Dapagliflozin Capsule, 10 mg (as the Non-SolvatedForm)

1. Fill empty capsule shells with Example 1 Part A stock granulation forcapsules (10.0% w/w as the non-solvated form), to provide 10 mgcapsules.

2. De-dust the capsules.

3. Weight sort the capsules.

The Example 1 (2.5 mg) and Example 2 (10 mg) capsules are used intreating obesity.

Example 3 Dapagliflozin Capsule, 100 mg (as the Non-Solvated Form)

Composition: 438.6 mg of dapagliflozin (Example 3 Part A) StockGranulation for Capsules (22.8% w/w as the non-solvated form), filled inGray, Opaque, Size #0 Capsule Shell.

A. Stock Granulation Composition

Ingredient Amount (% w/w) Dapagliflozin PGS¹ 22.8 Pregelatinized Starch,NF 15.0 Microcrystalline Cellulose, NF² 55.95 Sodium Starch Glycolate,NF 3.0 Silicon Dioxide, NF 2.0 Magnesium Stearate, NF³ 1.25 ¹This amountis expressed in terms of the amount of the dapagliflozin at 100% purity.The exact amount will vary depending on the purity of the dapagliflozin.²The amount of microcrystalline cellulose used will vary depending onthe purity of the dapagliflozin. ³The preferred amount is 1.25% (w/w).The range is 1.25-1.50% (w/w).

The stock granulation of Part A and the Example 3 capsules were preparedaccording to the following procedures.

B. Stock Granulation Procedure

1. Screen silicon dioxide.

2. Mix silicon dioxide with dapagliflozin in a suitable blender.

3. Screen the blend from Step 2, and mix again.

4. Screen pregelatinized starch and microcrystalline cellulose, ifnecessary.

5. Add ingredients form Step 4 to the blend from Step 3, and mix.

6. Screen sodium starch glycolate.

7. Add ingredient from Step 6 to the blend from Step 5, and mix.

8. Screen a portion of magnesium stearate.

9. Add ingredient from Step 8 to the blend from Step 7, and mix.

10. Densify the blend from Step 9.

11. Reduce the densified blend from Step 10.

12. Screen the remaining portion of magnesium stearate.

13. Add ingredient from Step 12 to the granulation from Step 11, andmix.

C. Example 3 Product: Dapagliflozin Capsule, 100 mg (as the Non-SolvatedForm)

1. Fill empty capsule shells with Example 3 stock granulation forcapsules (22.8% w/w as the non-solvated form).

2. De-dust the capsules.

3. Weight sort the capsules.

Examples 4 to 6

Tablets containing the SGLT2 inhibitor of formula Ia (dapagliflozin(S)-propylene glycol solvate (PGS) (or dapagliflozin PGS) were preparedin strengths of 2.5 mg (Example 4), 10 mg (Example 5) and 50 mg (Example6) as described below.

Example 4 Product: Dapagliflozin PGS Tablet, 2.5 mg A. TabletComposition

Ingredient Amount Dapagliflozin PGS¹ 3.075 mg MicrocrystallineCellulose, NF² 67.113 mg  Lactose Anhydrous, NF 25.000 mg  Crospovidone,NF 8.750 mg Croscarmellose Sodium, NF 3.750 mg Talc, USP 12.500 mg Silicon Dioxide, NF 2.875 mg Magnesium Stearate, NF³ 1.938 mg¹Dapagliflozin PGS is a propylene glycol solvate. The amount ofnon-solvated dapagliflozin is theoretically equivalent to 81.29% ofdapagliflozin PGS. The actual amount of dapagliflozin PGS will depend onthe “As Is” purity of the drug. ²This is the compensating excipient. Theamount used may vary depending on the “As Is” purity of the drug and/orthe actual amount of magnesium stearate used. ³The target amount is 1.94mg. Acceptable range is 1.55-2.33 mg.

The stock granulation of Part A and the Example 4 tablets were preparedaccording to the following procedures.

B. Stock Granulation Procedure

1. Deaggregate dapagliflozin PGS and magnesium stearate separately usinga suitable screen.

2. Mix dapagliflozin PGS with a portion of microcrystalline cellulose ina suitable mixer and transfer it into a suitable blender.

3. “Dry Rinse” the mixer used for mixing Step 2 with a portion ofmicrocrystalline cellulose.

4. Add the blend from Step 3 to the blend from Step 2.

5. Mix the mixture from Step 4 with remaining microcrystallinecellulose, portion of crospovidone, portion of croscarmellose sodium,portion of silicon dioxide and lactose anhydrous.

6. Add talc and intragranular magnesium stearate to the mixture fromStep 5 and mix.

7. Compact the powder blend from Step 6.

8. Reduce compact from Step 7 to form granules.

9. Mix the granules from Step 8 with remaining amounts of crospovidone,croscarmellose sodium and silicon dioxide.

10. Mix the granules from Step 9 with remaining amount of magnesiumstearate.

C. Example 4 Product: Dapagliflozin PGS Tablet, 2.5 mg

1. Setup the tabletting equipment.

2. Compress the Example 4 stock granulation into tablets (2.46% w/w),(2.5 mg).

Example 5 Product: Dapagliflozin PGS Tablet, 10 mg A. Tablet Composition

Ingredient Amount Dapagliflozin PGS¹ 12.300 mg MicrocrystallineCellulose, NF² 57.888 mg Lactose Anhydrous, NF 25.000 mg Crospovidone,NF  8.750 mg Croscarmellose Sodium, NF  3.750 mg Talc, USP 12.500 mgSilicon Dioxide, NF  2.875 mg Magnesium Stearate, NF³  1.938 mg¹Dapagliflozin PGS is a propylene glycol solvate. The amount ofnon-solvated dapagliflozin PGS is theoretically equivalent to 81.29% ofdapagliflozin PGS. The actual amount of dapagliflozin PGS will depend onthe “As Is” purity of the drug. ²This is the compensating excipient. Theamount used may vary depending on the “As Is” purity of the drug and/orthe actual amount of magnesium stearate used. ³The target amount is 1.94mg. Acceptable range is 1.55-2.33 mg.

The stock granulation of Part A and the Example 5 tablets were preparedaccording to the following procedures.

B. Stock Granulation Procedure

1. Deaggregate dapagliflozin PGS and magnesium stearate separately usinga suitable screen.

2. Mix microcrystalline cellulose, dapagliflozin PGS portion ofcrospovidone, portion of croscarmellose sodium, portion of silicondioxide and lactose anhydrous in a suitable blender.

3. Add talc and intragranular magnesium stearate to the mixture fromStep 2 and mix in a suitable blender.

4. Compact the powder blend from Step 3.

5. Reduce compact from Step 4 to four granules.

6. Mix the granules from Step 5 with remaining amounts of crospovidone,croscarmellose sodium and silicon dioxide.

7. Mix the granules from Step 6 with remaining amount of magnesiumstearate.

C. Example 5 Product: Dapagliflozin Propylene Glycol Solvate (PGS)Tablet, 10 mg

1. Setup the tabletting equipment.

2. Compress the Example 5 stock granulation into tablets (9.84% w/w).

Example 6 Product: Dapagliflozin PGS Tablet, 50 mg A. Tablet Composition

Ingredient Amount Dapagliflozin PGS¹ 61.660 mg MicrocrystallineCellulose, NF² 114.090 mg  Lactose Anhydrous, NF 62.600 mg Crospovidone,NF 21.910 mg Croscarmellose Sodium, NF  9.390 mg Talc, USP 31.300 mgSilicon Dioxide, NF  7.200 mg Magnesium Stearate, NF³  4.850 mg ¹Theamount shown is based on the amount of dapagliflozin PGS at 100% purity.The exact amount may vary depending on the “As Is” purity of thedapagliflozin PGS. ²This is the compensating excipient. The amount usedmay vary depending on the “As Is” purity of the drug and/or the actualamount of magnesium stearate used. ³The target amount is 4.85 mg.Acceptable range is 3.76-5.95 mg.

The stock granulation of Part A and the Example 6 tablets were preparedaccording to the following procedures.

B. Stock Granulation Procedure

1. Mix dapagliflozin PGS microcrystalline cellulose, lactose anhydrous,crospovidone, croscarmellose sodium, talc and silicon dioxide in asuitable blender.

2. Pass the mixture from Step 1 through a suitable mill.

3. Determine the yield from Step 1 and calculate the amount of magnesiumstearate required.

4. Mix the mixture from Step 2 in a suitable blender.

5. Mix the mixture from Step 4 with magnesium stearate.

6. Dry granulate the powder blend from Step 5.

7. Size the granulation from Step 6.

8. Determine the yield based on Step 7.

9. Mix the granules from Step 8 with remaining amount of crospovidone,croscarmellose sodium and silicon dioxide.

10. Mix the granules from Step 9 with remaining amount of magnesiumstearate.

C. Example 6 Product: Dapagliflozin PGS Tablet, 50 mg

1. Setup the tabletting equipment.

2. Compress the Example 6 stock granulation (19.7% w/w), into tablets(50 mg).

Example 7 SGLT2 Inhibitor Dapagliflozin PSG Increases Urinary Excretionof Glucose in Healthy Subjects

The glucosuric effects of dapagliflozin PGS results in significant lossof calories in the urine as compared to a known SGLT2 inhibitor (GSK869,682). As seen in FIG. 1, the results of an indirect comparison oftwo single ascending dose studies of SGLT2 inhibitors are shown. Thepanel at the top of FIG. 1 shows the amount of glucose excretion/daywithin 24 hours in healthy subjects taking 50, 100, 200 or 500 mg of GSK869,682. The panel at the bottom shows the amount of glucoseexcretion/day within 24 hours in healthy subjects taking 5, 20, 50 or100 mg of dapagliflozin PGS.

In a similar experiment, the effects of the C-arylglucosidedapagliflozin PGS and two O-arylglucosides on urine glucose excretion inhealthy individuals were compared. The results are shown in Table 4.

TABLE 4 Urinary Glucose Excretion over 24 hr in Healthy NormalVolunteers Following Administration of Selected SGLT2 Inhibitors GlucoseOutput Drug Dose over 24 hr Sergliflozin-A (O-  200 mg   12 gglucoside)*  500 mg   17 g AVE 2268 (O-glucoside) 1200 mg   14 g 2000 mg  21 g Dapagliflozin (C-glucoside)   5 mg ~32 g  20 mg ~64 g*Sergliflozin-A (GSK 869682) has the following structure

Example 8 SGLT2 Inhibitor Dapagliflozin PGS Increases Urinary Excretionof Glucose Over a 24 Hour Period in Patients with Diabetes

In a separate study, patients with diabetes were treated withdapagliflozin PGS at dose of 5, 25, or 100 mg or treated with placebo.The amount of urinary glucose excretion (g/day) was plotted as afunction of time and is shown in FIG. 2. The levels of urine glucose ofall subjects are shown in the plot and the mean urine glucose is in thedarker lines. Compared to subjects treated with placebo, subjectstreated with dapagliflozin PGS showed higher levels of urine glucoseexcretion.

Example 9 Patient Treated with SGLT2 Inhibitor Dapagliflozin for 12Weeks Exhibited Increased Urine Volume and Reduced Serum Uric AcidLevels

C-aryl glucoside SGLT2 inhibitors are known to reduce blood glucoselevels in diabetic patients, see U.S. Pat. No. 6,774,112, which isincorporated herein by reference in its entirety, but not known toincrease urine volume. In this experiment, the effects of dapagliflozinPGS on glucosuria and urine volume were examined. 47 drug-naïve ormetformin-treated type 2 diabetic patients with fasting serum glucoselevels no greater than 240 mg/dL were included in the study. Metforminreduces plasma glucose concentration by non-renal mechanisms and is notknown to induce glucosuria. The patients received various amounts ofdapagliflozin PGS or placebo on day zero. On day 1 and day 14, thepatients were examined for the levels of glucosuria and changes in urinevolumes. As shown in FIG. 3, urinary glucose levels increased within a24-hour time period in dapagliflozin-treated, but not inplacebo-treated, subjects. Within the dapagliflozin-treated group, theconstant rate of glucosuria over 24 hours was estimated to be about 2g/hr and 48 g/d (5 mg dapagliflozin PGS) and about 3 g/hr and 72 g/d (25mg and 100 mg dapagliflozin PGS). Dose-dependent reduction in fastingserum glucose (FSG) levels, and reduced postprandial glucose levels inoral glucose tolerance tests (OGTT) were also observed indapagliflozin-treated patients. Although both FSG and OGTT responsesimproved over a two-week period of time, no increase in urine volumeswas apparent during the same time period. Patients' serum uric acidlevels were not measured in this experiment.

Urine volume increases were observed in patients with prolongedtreatment of dapagliflozin PGS. In a separate experiment, diabeticpatients were treated with 2.5 mg, 5 mg, 10 mg, 20 mg, and 50 mgdapagliflozin PGS for 12 weeks (N=389, body mass index (BMI)>30, withglycosylated hemoglobin HbA1C levels around 7.7-8.0%). After 12 weeks oftreatment, the patient excreted 50-60 g glucose in urine every day, andshowed improved OGT and reduced HbA1C levels (0.71-0.9%, similar topatients treated with 1500 mg metformin, which showed HbA1C levels at0.73%).

Significantly, the patients treated with dapagliflozin PGS for 12 weeksshowed a 5-20% increase in urine volume. Further, patients receiveddapagliflozin PGS at all concentrations exhibit a decrease of serum uricacid levels after 12 weeks treatment. See Table 5. The average ofdecrease in serum uric acid levels was approximately 1 g/dL, whilepatients received placebo exhibited a reduction of serum uric acid byonly 0.16 mg/dL. Thus, dapagliflozin PGS decreased serum uric acidlevels and can be used as a treatment for hyperuricemia.

TABLE 5 Dapagliflozin PGS reduced serum uric acid in individuals after12 week- treatment dapagliflozin PGS (mg) 2.5 5 10 20 50 placebometformin Uric Acid −1.03 ± 0.81 −1.12 ± 0.84 −0.98 ± 0.66 −1.13 ± 0.78−1.14 ± 1.15 −0.16 ± 0.75 0.18 ± 0.53 (mg/dL) p-value vs. <0.0001<0.0001 <0.0001 <0.0001 <0.0001 placebo Values are means ± standarddeviation.

What is claimed is:
 1. A method for treating hyperuricemia in a mammalcomprising administering to the mammal in need of such treatment atherapeutically effective amount of a sodium glucose transporter 2(SGLT2) inhibitor in the absence of the administration of uric acidsynthesis inhibitors and uricosurics, wherein the SGLT2 inhibitor is

or a prodrug ester thereof or a propylene glycol solvate thereof.
 2. Themethod of claim 1, wherein the SGLT2 inhibitor is


3. The method of claim 1 wherein the SGLT2 inhibitor increases urinevolume and lowers serum total urate levels.
 4. The method of claim 1,wherein the mammal is a human.
 5. The method of claim 4, wherein thehuman has gout as a result of hyperuricemia, and the treatment ofhyperuricemia treats the gout in the human.
 6. The method of claim 1,wherein the SGLT2 inhibitor is administered before, after, orconcurrently with a supply of carbohydrate.
 7. The method of claim 1,wherein the SGLT2 inhibitor is administered in an amount sufficient totreat hyperuricemia without inducing hypoglycemia.
 8. A method fortreating hyperuricemia in a mammal comprising administering to themammal in need of such treatment a therapeutically effective amount of asodium glucose transporter 2 (SGLT2) inhibitor, wherein the SGLT2inhibitor is

or a prodrug ester thereof or a propylene glycol solvate thereof, andwherein the SGLT2 inhibitor is administered in an amount sufficient toremove about 1 mg/dL urate from the serum of the mammal.
 9. The methodof claim 8, wherein the mammal is a human.
 10. The method of claim 8,wherein the SGLT2 inhibitor is administered in an amount sufficient toincrease urine volume by 5-20%.
 11. The method of claim 8, wherein theSGLT2 inhibitor is


12. The method of claim 8 wherein the SGLT2 inhibitor increases urinevolume and lowers serum total urate levels.
 13. The method of claim 8,wherein the human has gout as a result of hyperuricemia, and thetreatment of hyperuricemia treats the gout in the human.
 14. The methodof claim 8, wherein the SGLT2 inhibitor is administered before, after,or concurrently with a supply of carbohydrate.
 15. The method of claim8, wherein the SGLT2 inhibitor is administered in an amount sufficientto treat hyperuricemia without inducing hypoglycemia.
 16. A method fortreating hyperuricemia in a mammal comprising administering to themammal in need of such treatment a therapeutically effective amount of asodium glucose transporter 2 (SGLT2) inhibitor, wherein the SGLT2inhibitor is

or a prodrug ester thereof or a propylene glycol solvate thereof, andwherein the SGLT2 inhibitor is administered in an amount sufficient toincrease urine volume by 5-20%.
 17. The method of claim 16, wherein themammal is a human.
 18. The method of claim 16, wherein the SGLT2inhibitor is


19. The method of claim 16 wherein the SGLT2 inhibitor increases urinevolume and lowers serum total urate levels.
 20. The method of claim 16,wherein the human has gout as a result of hyperuricemia, and thetreatment of hyperuricemia treats the gout in the human.
 21. The methodof claim 16, wherein the SGLT2 inhibitor is administered before, after,or concurrently with a supply of carbohydrate.
 22. The method of claim16, wherein the SGLT2 inhibitor is administered in an amount sufficientto treat hyperuricemia without inducing hypoglycemia.